A kind of preparation method and application of tyrosyl tRNA synthetase mutant
A technology of synthetase and tyrosyl, which is applied in the biological field, can solve the problems that have not yet been reported on the application of the preparation method of tyrosyl tRNA synthetase mutants, and achieve the effect of simple preparation method
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[0078] Materials and methods
[0079] Major equipment
[0080] (1) Ultra-clean workbench (Boxun Industrial Co., Ltd.)
[0081] (2) 5LBioflow3000 fermenter (NBS, USA)
[0082] (3) C25KC shaker (US NBS)
[0083] (4) UV spectrophotometer (BioRad)
[0084] (5) Freeze dryer (US FTS)
[0085] (6) AKTAexplore purification system (GEAmersham)
[0086] (7) High pressure homogenizer (PandaPlus2000 type, NiroSoavi, Italy)
[0087] (8) Microplate reader (perkinElmer1420, VICTOR3).
[0088] Material
[0089] (1) Strain and plasmid: Tmax highly expressed strain BL21 (F-ompThsdS(rB-mB-)galdcm, Novagen) and anti-Tmax monoclonal antibody were provided by aTyrPharma, USA;
[0090] (2) Purification filler: SP-SepharoseFastFlow; Sephadax-G50; Q-SepharoseFastFlow (Pharmaei);
[0091] (3) Protein building, yeast extract (Oxoid), casein hydrolyzate (Sigma);
[0092] (4) The rest are domestic analytically pure.
[0093] Media and common solutions
[0094] (1) Fermentation medium M9CA (1.0...
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